Unit 1.1.35.4 Soluble Nitrogen by The Kjeldahl Method

Ref: GLT.QA.AM.
1296
SABMiller plc – Group Technical
Page: 1 of 7
Analytical Method Version: 1.0
Version Date: 1.0
Title: Unit 1.1.35.4 Soluble Nitrogen By the Kjeldahl Method Review Date: 29/01/2019
Contents
1 Version & Revision Details.....................................................................................................2

1.1 Version 1 ......................................................................................................................2
2 Purpose and Principles ..........................................................................................................3
2.1 Purpose ........................................................................................................................3
2.2 Principles......................................................................................................................3
3 Scope ..................................................................................................................................... 3
4 References............................................................................................................................. 3
5 Definitions and Abbreviations.................................................................................................3
5.1 Definitions.....................................................................................................................3
5.2 Abbreviations................................................................................................................3
6 Method ................................................................................................................................... 3
6.1 Health and Safety.........................................................................................................3
6.2 Equipment ....................................................................................................................4
6.3 Reagents ......................................................................................................................4
6.4 Measurement Procedure..............................................................................................4
6.5 Reporting......................................................................................................................7
6.6 Verification....................................................................................................................7
POSITION NAME SIGNATURE DATE
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Title: Unit 1.1.35.4 Soluble Nitrogen By the Kjeldahl Version: Ref: GLT.QA.AM.1296
1.0 Page: 2 of 7
Method
1 Version & Revision Details
1.1 Version 1
This is the original document Version 1.

1.0 Page: 3 of 7
Method
2 Purpose and Principles
2.1 Purpose
This document describes a method to determine the nitrogen in wort derived from malt by the
Kjeldahl method.
2.2 Principles
Malt is first mashed to produce wort. The nitrogenous compounds in the wort are then digested
with hot sulphuric acid in the presence of a catalyst to give ammonium sulphate.
The digest is made alkaline with sodium hydroxide solution releasing ammonia which is distilled
into an excess of boric acid solution.
The ammonia is quantified by titration with standard acid solution.
3 Scope
This method can be applied to all worts prepared during the course of malt analysis.
4 References
Analytica-EBC Method 4.9.1: Soluble Nitrogen of Malt, Kjeldahl Method

Analytical Methods Unit 1.1.35.5 Total Nitrogen by the Kjeldahl Method
Analytical Methods Unit 1.1.45: Sampling of Barley and Malt
Analytical Methods Unit 1.1.16.1: Determination of Congress Wort Extract
5 Definitions and Abbreviations
Note: Section 5 is as specified in Analytical Methods Unit 1.1.35.5 Total Nitrogen by the
Kjeldahl Method
5.1 Definitions
5.2 Abbreviations
6 Method
6.1 Health and Safety
Note: Section 6.1 as specified in Analytical Methods Unit 1.1.35.5 Total Nitrogen by the
Kjeldahl Method
6.1.1 Hazards
6.1.2 Safety Requirements
6.1.3 Precautions
1.0 Page: 4 of 7
Method
6.2 Equipment
Note: Section 6.2 as specified in Analytical Methods Unit 1.1.35.5 Total Nitrogen by the
Kjeldahl Method
6.2.1 Equipment Requirements

6.2.2 Equipment Preparation
6.2.3 Maintenance
6.2.4 Calibration
6.2.5 Storage
6.3 Reagents
Note: Unless otherwise stated in the specific sub-sections Section 6.3 as specified in
Analytical Methods Unit 1.1.35.5 Total Nitrogen by the Kjeldahl Method
6.3.1 Reagents Required
a) As in Section 6.3 of Unit 1.1.35.5

b) Antifoam emulsion
6.3.2 Reagent Preparation

6.3.3 Standardisation
6.3.4 Disposal
6.4 Measurement Procedure
6.4.1 Sampling
Take a laboratory sample of malt as specified in Analytical Methods Unit 1.1.45
6.4.2 Sample Preparation
Prepare 50 ml Congress wort as specified in Analytical Methods Unit 1.1.16.1
6.4.3 Test Method
6.4.3.1 Test Method using distillation apparatus
a) Pipette 20 ml of wort into a 500 ml Kjeldahl flask.

b) Add 2 to 3 ml of concentrated sulphuric acid.
c) If necessary, add antifoam to prevent excess foaming.
d) Gently evaporate, almost to dryness, with minimal charring.
e) Add about 10 g of powdered catalyst mixture (or tablets) to the dried sample and mix
thoroughly.
f) Carefully add 20 ml 98% H2SO4 to the flask and swirl gently to mix and thoroughly wet the
contents.
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Method
g) Allow to digest until frothing stops.

h) Apply heat to boil the digest briskly until the brown colour disappears.
i) Continue heating for 30 min.
Note: It is most important that direct heat does not come into contact with the flask above
the level of the liquid and that the vapour should be seen to be refluxing in the lower
regions of the neck of the flask.
j) Allow the digest to cool.
Note: If the contents solidify, excessive acid has been lost during digestion and ammonium
sulphate could have been volatilised. Should this occur the digestion should be repeated.
k) Carefully dilute the digest with 250 ml purified water.

l) Add anti-bumping agent then slowly add about 70 ml NaOH solution, so that two distinct
layers are formed.
m) Add 25 ml 20 g/l boric acid solution and 0.5 ml screened indicator to the collection flask and
swirl to mix.
n) Without disturbing the layers, fit the trap and connect to the condenser unit.
o) Ensure that the exit tube from the condenser dips below the surface of the boric acid
solution in the collection flask.
p) Turn on the heaters.
Note: The heaters should be switched on before connecting the flask to minimize the
danger of liquid passing back through the condenser.
q) Swirl the contents of the flask to ensure rapid mixing and apply full heat.
Note: Immediately after mixing, it is advisable that the flask containing the boric acid is
withdrawn momentarily, to let the exit tube drain and to equalize the pressure in the
distillation flask.
r) Collect about 180 ml of distillate.

s) Using a 25 ml or 50 ml burette titrate the ammonia with the standard acid to the grey end
point.
t) Record the amount of acid used (ml).
Blank reaction
a) Weigh 1.000 g sucrose into a tared, completely dry Kjeldahl flask.

b) Proceed as per this Section, instructions e) to t).
6.4.3.2 Test Method using Buchi Furnace and Buchi Distillation Apparatus
a) Pipette 20 ml of wort into a 500 ml Kjeldahl flask.

b) Add 2 ml of concentrated sulphuric acid.
c) If necessary, add antifoam to prevent excess foaming.
d) Gently evaporate, almost to dryness, with minimal charring.
e) Add about 10 g of powdered catalyst mixture (or tablets) to the dried sample and mix
thoroughly.
1.0 Page: 6 of 7
Method
f) Carefully add 18 ml 98% H2SO4 to the flask and swirl gently to mix and thoroughly wet the
contents.
g) Allow to digest until frothing stops.
h) Next step is the mineralisation process in Buchi furnace which should continue until the
brown colour changes into green
i) Continue heating for about 30 min.
j) Dose the digest with 50 ml purified water and 70 ml NaOH solution
k) Insert the Erlenmeyer containing 25 ml 20 g/l boric acid solution and about 8 drops of
Tashiro indicator and turn on distillation for 4 minutes.
l) Collect about 150 ml of distillate.
m) Using stand-alone titrator with stirrer titrate the ammonia with the standard 0.1 N sulphuric
acid to the purple end point.
6.4.4 Calculations
The soluble nitrogen content is calculated either per litre of wort, or as a percentage of dry malt, or
of the total nitrogen content (Kolbach Index).
Soluble nitrogen content litre of wort:
NV (mg/l) = T x 14 x 100
V
Where:
NV = soluble nitrogen content in mg/litre
T = ml of 0.1M acid required to neutralize ammonia after subtracting the reagent blank
V = volume of sample taken (ml)
Soluble nitrogen content as a percentage of dry malt:
NS (%) = NV x E 1
10000 x EW
Where:
NV = soluble nitrogen content in mg/litre
E1 = extract content on dry malt in % (m/m)
EW = g of extract in wort in g/100 ml of wort
10000 = correction factor to obtain the result as a percentage
Soluble nitrogen content as a percentage of the total nitrogen content (Kolbach Index):
NK = NS x 100
N
Where:
NK = soluble nitrogen content as a percentage of total nitrogen in % (m/m)
NS = soluble nitrogen content on dry malt in % (m/m)
N = total nitrogen content on dry malt in % (m/m)
1.0 Page: 7 of 7
Method
6.5 Reporting
Soluble nitrogen per litre of wort: Express the result as mg/litre to the nearest whole number.
Soluble nitrogen as a percentage of dry malt: Express the result as % (m/m) to 2 decimal places.
Soluble nitrogen content as a percentage of the total nitrogen content (Kolbach Index): Express the
result as % (m/m) to the nearest whole number.
6.6 Verification
Analytical Methods Unit 0 Verification Routines

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Ref: GLT.QA.AM.

Analytical Method Version: 1.0

Version Date: 1.0

1 Version & Revision Details.....................................................................................................2

POSITION NAME SIGNATURE DATE

1 Version & Revision Details

This is the original document Version 1.

2 Purpose and Principles

Analytica-EBC Method 4.9.1: Soluble Nitrogen of Malt, Kjeldahl Method

5 Definitions and Abbreviations

6.1 Health and Safety

6.2.1 Equipment Requirements

6.3.1 Reagents Required

a) As in Section 6.3 of Unit 1.1.35.5

6.3.2 Reagent Preparation

6.4 Measurement Procedure

Take a laboratory sample of malt as specified in Analytical Methods Unit 1.1.45

6.4.2 Sample Preparation

Prepare 50 ml Congress wort as specified in Analytical Methods Unit 1.1.16.1

6.4.3 Test Method

6.4.3.1 Test Method using distillation apparatus

a) Pipette 20 ml of wort into a 500 ml Kjeldahl flask.

g) Allow to digest until frothing stops.

k) Carefully dilute the digest with 250 ml purified water.

r) Collect about 180 ml of distillate.

a) Weigh 1.000 g sucrose into a tared, completely dry Kjeldahl flask.

a) Pipette 20 ml of wort into a 500 ml Kjeldahl flask.

Soluble nitrogen content litre of wort:

Soluble nitrogen content as a percentage of dry malt:

Analytical Methods Unit 0 Verification Routines

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